CN1521275A - Multiple arrays with surface energy transition to maintain separation of samples on the arrays - Google Patents

Abstract

An array unit comprising a substrate surface carrying multiple chemical arrays each comprising multiple features and each array being surrounded by a surface energy transition to maintain a separation between separate liquid samples applied to the arrays, and wherein the substrate surface is physically uninterrupted over a continuous region carrying the arrays. Methods of array fabrication and use are further provided.

Description

Has the surface energy saltation zone to keep a plurality of arrays of sample separation on the array

Technical field

The present invention relates to the array such as polynucleotide array (for example DNA array), they have good application at aspects such as diagnosis, screening, gene expression analysis.

Background technology

In discussion below and the application's the full text, though quoted a lot of reference, these documents of quoting all can not constitute the application's prior art.

Known number of chemical array, for example polynucleotide array or protein array (for example DNA or RNA array), they have been used as such as diagnosis or screening implement.The polynucleotide array comprises and comprises the not zone of homotactic polynucleotide usually that these zones are arranged on the substrate in predetermined layout (configuration) mode.These zones (being sometimes referred to as " feature ") are positioned at on-chip position separately (" address ") and locate.When array is subjected to time spent of doing of sample, will show observable in conjunction with pattern (pattern).When array is read, thisly can be detected in conjunction with pattern.For example, all the polymerized nucleoside acid targets (for example DNA) in the sample can carry out mark by suitable marker (for example fluorescent chemicals), and the phosphor pattern on the array can accurately be observed after it is subjected to sample effect.Suppose that not homotactic polynucleotide is all correctly deposited the so viewed existence and/or the concentration that will show one or more polynucleotide compositions in the sample in conjunction with pattern according to predetermined layout.

The biomacromolecule array can deposit on the substrate or by in-situ synthetic method by the biomacromolecule (for example by synthetic or natural origin) that will obtain in advance and create.The method of the biomacromolecule that deposition is obtained comprises: in its pack into pin or kapillary, described pin or kapillary are contacted with the surface, for example US 5,807, described in 522, perhaps deposit by ejecting from the pulse sprinklers such as ink gun, for example PCT discloses among text WO 95/25116 and the WO98/41531 and elsewhere is described.Can think that this type of deposition method forms each feature (that is to say that each feature has only a circulation, during this period, the previous biomacromolecule that obtains is fixed on the substrate) by a fixed cycles.In position among the preparation method, a lot of different reagent droplet are deposited on the given target site, so that form the final feature probe of described feature (thereby on the substrate of array, synthesized) by pulse sprinklers or other devices.In-situ preparation method comprises: US 5,449, the method for the synthetic peptide array of describing in 754, US 6,180,351 and WO98/41531 and the reference quoted thereof described in the method for synthetic polynucleotide, also can use the pulse sprinklers deposited reagent.The in-situ method of preparation polynucleotide array adopts identical tradition to repeat operation at a plurality of different each places, address that remains in the address of this formation feature usually, and described traditional operation that repeats is used to form the polynucleotide feature by known chemical means by nucleoside agents on carrier.At each feature place to be formed is arranged, this repetition operation can be counted as a plurality of in the fixed cycles of back: (a) in first repeats, selected activatory nucleosides (monomeric unit) is coupled on the functionalized upholder by the phosphorous acid ester bond, perhaps in repetition subsequently, it is coupled to and is combined on the on-chip nucleosides (just nucleosides modify substrate); (b) alternatively, the unreacted hydroxyl that is combined in on-chip nucleosides can be closed (being also referred to as " adding cap " sometimes); (c) with the phosphorous acid ester bond oxidation of step (a) to form the phosphoric acid ester bond; And (d) blocking group is removed (go protection) from up-to-date being combined in the on-chip nucleosides of institute's link coupled step (a), so that be the generation of the circulation next time avtive spot in these steps.Can deposit on the expection feature locations specific on the array by the drop that will contain activator and phosphoramidite (phosphoramidite) and carry out coupling.Provide final going to protect step, in this step, nitrogenous base and phosphate-based going are simultaneously protected by under known condition, handling with ammonium hydroxide and/or methylamine.Add cap, oxidation and go to protect and to realize by the monoblock substrate being handled (" submergence ") with the suitable reagent of one deck.Functionalized upholder (first circulation in) or de-protected coupling nucleosides (in the follow-up circulation) provide the link group for being combined in on-chip part (moiety), so as with will in step (a), carry out the next nucleosides formation of link coupled phosphorous acid ester bond.Nucleoside base is final go protection can by in another submergence program, utilize in known manner alkaline condition for example ammonium hydroxide realize.Usually select for use a pulse sprinklers or other dividers to deposit a kind of monomeric unit.

Described above-mentioned polynucleotide synthetic chemical mechanism in the following document in detail, for example, Caruthers, Science 230:281-285,1985; Itakura et al., Ann.Rev.Biochem.53:323-356; Hunkapillar et al., Nature 310:105-110,1984; And " Synthesis ofOligonucleotide Derivatives in Design and Targeted Reaction ofOligonucleotide Derivatives ", CRC Press, Boca Raton, Fla., pages 100 et seq., US 4,458,066, US 4,500,707, US 5,153,3 19, US 5,869,643, EP 0294196 etc.Phosphoramidite and tris phosphite method are used the most extensively, but also have some other method, comprise phosphodiester method, phosphotriester method and H-phosphonic acid ester method.Substrate functionalised usually so that combine with sedimentary first monomer.For example, at US 6,258,454 and Southem, E.M., Maskos, U.and Elder, J.K., Genomics, 13,1007-1017 has described suitable technique in 1992, is used to utilize this type of link part to make substrate functionalized.In the preparation of array, in any one circulation, different monomers and activator can be deposited on the different positions of substrate, thereby make the different characteristics of the array after finishing have different desired biomacromolecule sequences.In each circulation, one or more intermediate steps be may also need, for example conventional oxidation in the in-situ preparing of polynucleotide array, cap and rinse step (similarly, these steps can be carried out) added in the submergence program.

US 6,242,266, and US 6,232,072, and US 6,180,351 and US 6,171,797 in the further details that biomacromolecule by depositing previous acquisition or in-situ method prepare the biomacromolecule array is disclosed.United States Patent (USP) 6,319 discloses useful especially link-group compound and method in 674 and 6,444,268.A kind of device also is provided in these patents, can have changed the surface energy of substrate, thereby in the array preparation process, controlled sprawling of deposition drop by this device.

In the preparation process of array, the amount of available polynucleotide often seldom, and is and very expensive.In addition, the sample size that can be used to test often also seldom.Under these conditions, the array that needs the less and tight distribution of use characteristic.For example, can have thousands of features on the array.But the array (for example having only 100 features) that has in some cases, the feature of much less is just enough.Have still less that a plurality of arrays of feature can be accommodated on the same substrate, and with different sample effects.In this case, in order to stop the mixing of different samples, can set up physical barrier between different arrays, liner or other device for example is applied to a sample liquids on the array and contacts with sample liquids on being applied to adjacent array so that stop.This may be before applying sample liquids the such gasket construction of assembled by hand, produce unexpected array contact or pollute the chance of (for example contacting the finger of array) thereby provide, cause from very expensive array, can't obtaining significative results from the user.Though can before the preparation array, obstruct be fixed on the substrate, possible obstruct breaks away from physical interference (for example when deposition head need be close to substrate surface and move) to preparation process and the treating processes but this may cause, and needs extra preparation process.

The present invention recognizes preferably can provide a plurality of arrays on substrate, it can act on different liquid samples (or different piece of same sample), provide a kind of device to stop the contact between different samples or the sample part simultaneously, this device does not require complicated processing and is easy to and makes.

Summary of the invention

The invention provides a kind of array element, comprising a plurality of chemical array (biological example macromole or other polymer arrays) that is positioned on the substrate surface.Each array comprises a plurality of features, and each array all surrounds by the surface energy saltation zone, to keep the separation between the isolating liquid sample that is applied on the array.Substrate surface is physically continual on the successive zone of carrying array.

The present invention also provides the method for preparing array element of the present invention.This method can comprise to substrate surface has the site deposition of feature to be formed to contain the drop of biomacromolecule precursor, thereby makes precursor combine with the surface in described site.Can repeat this operation in each described site, the precursor that has before connected is combined with next step sedimentary precursor, be produced out up to array.In the method, after repeating aforesaid drop deposition, compare with zone between array, the regional hydrophobicity that forms array becomes littler.The present invention also provides a kind of device, can carry out the method for preparing array element according to of the present invention, the present invention also provides a kind of computer program that carries computer-readable code, when it is carried out by computer, can carry out preparation method of the present invention (for example on aforesaid device).

The present invention also provides the method for using array element, and described array element comprises a plurality of biomacromolecule arrays that are positioned at substrate surface, and each array all comprises a plurality of features, and array by physically between continual array surf zone separate.This method comprises apply isolating liquid sample on each array, thereby makes a plurality of same times of isolating sample be present in substrate surface, keeps separate stage by zone between array simultaneously.For example, this method can be applied to array of the present invention, thus the separation of the sample room that applies that made between the stronger array of hydrophobicity zone maintenance.

The another kind of method of use provided by the present invention array element of the present invention comprises reads the code relevant with array element, and from described code or with file that described code is linked the relevant information that should be applied to the liquid sample volume on the array of acquisition.

Any or multiple in the benefit that all respects of the present invention can provide following or other are useful.For example, can on substrate, provide a plurality of arrays, can act on different liquid samples (or different piece of same sample).Simultaneously, the contact between different samples or the sample part can be suppressed, but this and do not require complicated processing, and be easy to make

Description of drawings

Fig. 1 shows the array component of carrying such as a plurality of arrays that can make by method of the present invention;

Fig. 2 is the enlarged view of the part among Fig. 1, shows the feature of a plurality of ideal points or array;

Fig. 3 is the enlarged view of the part among Fig. 2;

Fig. 4 shows the droplet dia of different volumes static state (fixing) drop on the array that is positioned at array element of the present invention, and it is the function of contact angle;

Fig. 5 shows the liquid volume that caused by the surface energy saltation zone to long-pending discontinuous of liquid contact surface, and the surface energy saltation zone is since on the unit of the present invention the hydrophobicity of array cause less than zone between array;

Fig. 6 shows the apparatus and method of use provided by the present invention array element of the present invention;

Fig. 7 shows the use of array element of the present invention; And

Fig. 8 shows the another kind of array layout of array element of the present invention.

For the ease of understanding, in feasible, used same label to mark identical element total in the different accompanying drawings.The different members (for example array 12a, 12b, 12c can usually be called " array 12 ") in the big class pointed out in the different letters that are positioned at the same numbers back.Accompanying drawing is not necessarily pro rata.

Embodiment

In the present invention, unless opposite purpose occurred, following term all refers to specified feature." biomacromolecule " is the polymkeric substance that repeating unit constituted by one or more types.Biomacromolecule can be found in biosystem usually, comprise that specifically polysaccharide (for example carbohydrate), peptide are (when using this term, comprise with/protein and the polypeptide that are not connected with polysaccharide) and polynucleotide, and their analogue, such as the compound that constitutes or contain following material by following material: amino acid analogue or non-amino acid group, perhaps nucleotide analog or non-nucleotide group.This comprises that its conventional skeleton has been replaced by the polynucleotide of non-natural or synthetic skeleton, and one or more conventional base has been replaced by the nucleic acid (or synthetic or natural analog) of the group (natural or synthetic) that can participate in Watson-Crick type hydrogen bond action.Polynucleotide comprises strand or multichain conformation, and wherein one or more in these chains can match fully or can not match fully with other chains." Nucleotide " is meant the subunit of nucleic acid, have phosphate-based, five-carbon sugar and nitrogenous base, and the functional analogue of this type of subunit (synthetic or natural), wherein the described analogue of polymer form (for example polynucleotide) can be hybridized in sequence-specific mode mutually with natural polynucleotide, and the hybridization mode of its hybridization mode and two kinds of natural polynucleotides is similar.For example, " biomacromolecule " comprises DNA (comprising cDNA), RNA, oligonucleotide and PNA, and US 5,948,902 and citing document (by reference its full content being included in herein) at this described in other polynucleotides, no matter its source is how." oligonucleotide " is meant that generally length is about the Nucleotide polymer of 10 to 100 Nucleotide, and " polynucleotide " comprises the Nucleotide polymer that contains any number Nucleotide." biomonomer " is meant one unit; can be linked to form biomacromolecule (for example have two single amino acids or Nucleotide that link group, one or two in the wherein said linking group can have removable blocking group) with identical or other biomonomer." biomacromolecule precursor " is the more junior unit of biomacromolecule, can connect to form biomacromolecule tail by the chemical bond tail.Biomonomer is one type a biomacromolecule precursor, but the biomacromolecule precursor can comprise two or more the monomeric unit that is linked.Fluid is meant liquid (for example, the solution of biomacromolecule or biomacromolecule precursor).

" array ", unless opposite purpose occurred, the any one dimension, two dimension or the three dimensional arrangement that comprise the addressable area that has the particular chemical part, wherein, described chemical part (for example being positioned at the not homotactic biomacromolecule of having of different zones, as polynucleotide) is associated with this zone.When substrate was vesicular structure, each zone can be extended to the third dimension; And when substrate is non-porous structure, then do not have the measuring of the third dimension (thickness) of any essence.Array is " addressable ", because it comprises a plurality of zones of containing different piece (for example different polymerized nucleoside acid sequences), thereby make the zone (" feature " of array or " point ") that is positioned at predetermined site place specific on the array (" address ") will detect a concrete target or a class target (though feature also might unexpectedly detect the target that does not correspond to this feature).Array features generally is a homogeneous, and feature is spaced apart the district usually and separates, but not necessarily leaves no choice but like this.For array, " target " is meant the part of mobile phase (normally fluid) lining, and the probe that combines with substrate (" target-probe ") that it awaits being positioned at each zone detects.But " target " or " target-probe " can be with by another evaluation that (therefore, any one can be the unknown mixture that contains polynucleotide, awaits by evaluated with combining of another)." array layout " or " array characteristic " can exchange use, be used in reference to one or more physics for array, chemical or biological characteristic, for example the layout of array internal feature and array are in on-chip layout, one or more characteristic dimensions, perhaps in some indication of the person's character of given feature site part or function (for example chemistry or biological), perhaps how array should (for example be operated, the condition of array and sample effect, perhaps array is read specification sheets, perhaps at sample effect afterwards from the expection signal or the range of signal of array controlling features).For polynucleotide, " hybridization " and " combination " can exchange use." controlling features " on the array is meant those features of checking array or sample performance during using array.For example, controlling features can be negative control (behind sample effect, the expection signal seldom or do not have) or positive regulation (behind sample effect, expection signal higher).

" plastics " are meant the synthetic organic polymer of any high molecular (for example at least 1,000 gram/mole, even at least 10,000 or 100,000 gram/moles).

When " flexibility " is used to describe substrate or substrate thin slice (web), be meant that substrate can be around the roller double flat of radius less than 1.25cm.Substrate can repeat bending like this and stretch at least 100 times and can not produce inefficacys (for example ftracture) or viscous deformation on either direction.This bending must be within the elastic limit of material.Aforesaid flexible test carries out under 20 ℃.

" thin slice " is meant the long continuous substrate sheet material of growing up in wide.For example, the long-width ratio of thin slice can be at least 5/1,10/1,50/1,100/1,200/1 or 500/1, even at least 1000/1.

When a project is called as " long-range " with respect to another project, be meant that these two projects are arranged in different buildingss at least, can be separated by 1 mile, 10 miles or at least 100 miles at least." interchange " information is meant that the data that will represent this information transmit by suitable communication passage (for example individual or public network) with the form of electrical signal." forwarding " project is meant this project is gone to another place from the three unities, can shift this project by physical means or other means (under the possible situation), comprise: at least for data, the medium that will carry data by physical means shifts or exchanges data.Array " assembly " can only be array with substrate, wherein array deposition is on this substrate, though assembly also can be the form (outer cover that for example has chamber) comprising the encapsulant of other features." chamber " is meant the volume (though can pass through one or more port access chambers) of sealing.It is to be further appreciated that in the application's full text, use term " preceding ", " back ", " top ", " on " and during D score, only adopted its relative meaning.When mentioning the project of odd number, also comprise the possibility that has a plurality of same projects." can " be meant selectable.Any method that is described in detail can given order be implemented when describing in detail, perhaps also can carry out according to other feasible in logic orders.

" pulse sprinklers " is meant any device that can distribute drop in the forming process of array.Pulse sprinklers when work, to being close to the exit or the liquid of nozzle gives a pressure pulse (for example by piezoelectricity or thermoelectric element), thereby make drop to distribute from described outlet or through hole.

For example, the thickness of " linking layer " that is connected with the surface can be less than 200 dusts, even less than 10 dusts (or thickness is less than 8,6 or 4 dusts).The minimum binding affinity of polynucleotide, protein, nucleosides or the amino acid of layer like this can be 10 ⁴To 10 ⁶Unit/μ ²If desired, can utilize the thickness of UV or X ray elliptical polarizer evaluation layer.

When " physically continual " when being used to describe substrate surface, being meant does not have physical barrier, and described physical barrier can diffuse out this array to stop it by receiving fluids.The size of physical barrier is enough greatly to stop the liquid-flow between array, but it is and different with the obstruct of receiving fluids, but the obstruct of described receiving fluids be since the surface energy that causes of chemical constitution discontinuous produce (for example, hydrophobic region and hydrophobicity littler face the zone mutually at the interface).For example, physically continual surface can be for example do not surround array wall and so on extend to the above surface that surpasses the physical barrier of 10 microns (or above 5,2 or 1 microns) of substrate surface.Lip-deep " zone " is the surface portion of the adjacency that is connected.That intra-zone can have is discontinuous (for example, the zone interconnects between array, but but there is the zone of load characteristic in portion within it, and the zone of these load characteristics is not the part in zone between array)." continuously " zone is continual (that is to say, extend fully between its outside dimension).Therefore, the successive zone of load characteristic is the sequential portion (so also comprising zone between feature) of the substrate of any object between load characteristic and the feature.When the successive zone of load characteristic is called as physically continual the time, then this is meant between the outer boundary of the surface portion that has array in it does not have physical barrier.

" group " relevant with chemical structural formula comprises the group that replaces and do not replace form.Substituent can comprise that low alkyl group, halogen or other can not produce the interferential substituent to expected results substantially.

" low alkyl group " is the alkyl that contains 1 to 6 C atom, can only have any in 1,2,3 or 4 C atom.

At US 6,444, provided the definition of " surface energy " in 268.

Liquid and the surface " contact angle " be meant from the teeth outwards the drop border and this surface between measured acute angle.The measurement of contact angle is known, can realize by multiple instrument, for example can be from First Ten Angstroms, and Portsmouth, VA, the FTA200 that U.S.A. obtains.Water or the waterborne liquid contact angle ratio on the stronger surface of hydrophobicity (surface energy is higher) is the littler lip-deep contact angle of hydrophobicity big (for example, the contact angle of water droplet is spent greater than 50 on the hydrophobic surface, even spends greater than 90).The contact angle of array (being sometimes referred to as " average contact angle " or " effectively contact angle ") is meant the average contact angle in zone between the feature of this array and feature.Unless stated otherwise, otherwise contact angle all is with aquametry.

" link agent density " or " adding cap agent density ", be meant unit surface internal chaining molecule or add the number of cap molecule.When determining link agent density, the link agent is counted, and no matter whether they link to each other with probe or self is added cap.For adding cap agent density, have only in adding of directly linking to each other with substrate surface, the cap agent was calculated in.Do the time spent if having the different zones of the substrate surface that homogeneous forms is subjected to identical component under identical condition link agent, the link agent density in these zones will be considered to " identical " so, wherein, described link agent combines with the surface with identical density.

" probe density " is a kind of abbreviation, is meant the number that links molecule or probe molecule in a feature on the unit surface.This term can exchange with " feature probe density " and use, and has same implication.Therefore, when determining probe density, the zone is not all considered between any feature that is substantially free of probe.Thereby " probe density " in a zone is different with characteristic density (number of features in the unit surface), and separate.

The step of any method herein can given order be implemented when describing in detail, perhaps also can carry out according to other feasible in logic orders.Whole patents of being quoted among the application and other reference, all by reference it is included in herein at this, but, except the inconsistent content (comprising definition) of any and the application in these patents or the reference (being as the criterion with the application in this case).

In array element of the present invention, the surface energy saltation zone can be provided by surf zone between array, surf zone is between the zone of carrying array between described array, and surrounds the zone of described carrying array, and stronger than the hydrophobicity of besieged zone iimit.In a kind of arrangement mode, the border in aforementioned besieged zone is the border of array (that is to say zone next-door neighbour array boundary between array).In a kind of structure, before the zone between array, the volume of the water that array can hold can reach 1mL, can reach 0.4mL usually, more commonly reaches 0.1mL at water intrusion.In another kind of layout, the volume of the water that array features can hold can reach V, and wherein V is provided by following formula:

$V(r,{\mathrm{\θ}}_c)=\frac{1}{3}\mathrm{\π}{\left(\frac{r}{\cos (\frac{\mathrm{\π}}{2}-{\mathrm{\θ}}_c)}\right)}^{\frac{1}{3}}(2-3\sin (\frac{\mathrm{\π}}{2}-{\mathrm{\θ}}_c)+\sin {(\frac{\mathrm{\π}}{2}-{\mathrm{\θ}}_c)}^3)$

Wherein, V (r, θ _c) be the volume of sessile drops; R is the radius of circular array, and perhaps when array is not circle, r is the radius of the circle that equates with array area; θ _cBe contact angle, spend (perhaps greater than 30,40,50 or 60 degree) and can spend (or less than 70 or 65 degree) less than 80 greater than 20.Perhaps, θ _cCan be contact angle greater than intermediate value, described intermediate value in above-mentioned angle any one and below between the array listed between any one in the more wide-angle in zone (for example between 80 degree and 90 degree).

The contact angle of array features can be less than 20 degree (or less than 15,10 or 5 degree), and the feature total area can account at least 30% (or at least 40%, 50%, 60% or at least 80%) of each array area.The average contact angle of array itself can be greater than 20 degree (perhaps greater than 30,40,50 or 60 degree) and less than 80 degree (or less than 70 or 65 degree), and the contact angle in zone can be greater than 80 degree (even greater than 90,95,100,105,110,115,120 or 130 degree) between array.The shape of array can be " circle ".In this linguistic context, " circle " only means and is not that trilateral neither rectangle, and can comprise oval-shaped array.Minimum interval between the adjacent array can be at least 2 times of mean distance between the feature of array, or is 3,4,5 or 8 times of this mean distance at least.And 10 (perhaps 9 or 5) that the minimum interval between the adjacent array can be not more than mean distance between the array features doubly.

Surf zone can comprise the dissimilar molecule that is connected with substrate surface between array.Each array features also can comprise the dissimilar molecule that is connected with substrate surface, and has only a subclass of described dissimilar molecules that biomacromolecule is covalently bound on the inner surface of feature.For example, described dissimilar molecule can be US 6,444, first and second silane described in 268, by reference this patent is included in herein herein, and in this case, described subclass will be second silane (first silane does not combine with any biomacromolecule on the array element).

In each array, array needs not to be (that is to say, equate at interval between the feature and have an identical size) of homogeneous.For example, each array can have interior region and surround the external region of interior region, and a plurality of features are contained in each zone, and wherein, the ratio of the shared external region of feature area is bigger than the ratio of the shared interior region area of feature.Thereby this external region can provide the hydrophilic/hydrophobic border between array and the array interior region.Array in the external region can have same composition.

As previously mentioned, in the method for the above-mentioned array of use of the present invention, isolating liquid sample (can be the different piece of different samples or same sample) can be placed on the one array, and keeps separate stage each other.In array of the present invention, the separate stage between the sample that is applied can be kept in zone (with respect to the littler array region of hydrophobicity) between the stronger array of hydrophobicity.Thereby, though people can further provide physical barrier between array, between array, can not have this type of physical barrier in the method for the invention, thereby avoided possible processing and pollution problem.In described method, array can keep in touch 1 hour, 5 hours or 10 hours with the isolating liquid sample that is applied on this array at least at least at least.For example, in the polymerized nucleoside acid hybridization, at elevated temperatures, sample can keep in touch about 12 hours with array, and this is known for the polynucleotide hybridization array.

In the method for preparing array element of the present invention, substrate surface can comprise the composition that combines with substrate surface, and described composition had both covered zone between array, also covers the zone that forms array, comprises the feature site.When deposition contained the drop of biomacromolecule precursor, at least a composition in the described composition can be with described precursor combination from the teeth outwards.Above-mentioned composition can comprise the dissimilar molecule that is connected with substrate surface, and has only a subclass of described dissimilar molecules biomacromolecule can be covalently bound on the inner surface of feature.For example, substrate surface can be by US6, and the hydrophobic mixture of first and second silane described in 444,268 institute applies fully, and the biomacromolecule feature is formed on on-chip different arrays place.Deposition contain biomacromolecule precursor (biological example monomer) drop first the circulation in, precursor only is bonded to the feature place by second silane.After a plurality of circulations, the hydrophobicity in feature site reduces, thereby the feasible hydrophobicity that forms the zone of array also becomes littler than zone between array.

Computer program of the present invention can comprise computer-readable storage media, is wherein storing computer program, and described computer program can be carried out method described herein by control device.For example, any computer-readable recording medium that is used for any purposes herein can comprise: light or magneticstorage (for example fixed or portable disk or other devices), perhaps solid-state memory.

With reference now to Fig. 1-Fig. 3,, array component 15 of the present invention can comprise substrate, for example the form of this substrate can be rigid substrate 10 (for example transparent non-porous material of finite length, for example glass or silica), carrying one or more arrays 12 (for example array 12a, 12b, 12c) above, these arrays distribute along the smooth upper surface 11a of substrate 10, and all arrays are all separated by surf zone 14 between same array, and surf zone 14 surrounds each array in the array 12 between array.Surf zone can be considered to just extend beyond the outermost of array 12 among Fig. 1 between array.The successive zone of carrying array 12 comprises zone 14 between array 12 and array.Substrate 10 also can be flexible.Each array 12 all and array on the surperficial 11a that extend in the common space, occupied himself zone (so these zones do not extend between array in the zone 14).The back side 11b of substrate 10 does not carry any array 12.Can the array on the substrate 10 be designed,, can be: specimen to be used to test the sample of any kind; Reference sample; The combination of above-mentioned sample; The perhaps known mixture (in this case, may comprise the feature of carrying unknown nucleotide sequence to be assessed in the array) of polynucleotide, protein, polysaccharide etc.Though 30 arrays 12 have been shown among Fig. 1, should be appreciated that in the embodiment of substrate 10 and use substrate 10, can use the array 12 of desired any number, for example at least 5,10,30,50,100, perhaps at least 200.According to application target, any one in the array 12 or all can be identical or differ from one another, and each all contains a plurality of points or feature 16 that biomacromolecule constituted by the polynucleotide form.In illustrated embodiment, the shape of array 12 roughly is (but also may be other shape, for example substantially oblong-shaped) of circle.

Typical array 12 can contain more than 5,10,20,30,100 even 150 features at least.For example, the width of feature (for the point of circle, just diameter) can be at 10 μ m in the 1.0cm scope.In other embodiment, the width of each feature can be at 1.0 μ m in the 1.0mm scope, and comparatively commonly 5.0 μ m are to 500 μ m, and more commonly 10 μ m are to 200 μ m.The areal extent of non-circular character can be equivalent to have the areal extent of the circular feature of aforementioned width (diameter) scope.At least a portion or all feature have different compositions (for example, when each feature with same composition any repeats to be excluded, remaining feature can account for the total characteristic number at least 5%, 10% or 20%).

The area of each array 12 can be less than 400mm ², even less than 200mm ², 100mm ², or less than 50mm ²Or 20mm ²In a lot of embodiment, particularly when substrate 10 was rigidity, its shape can be generally rectangular solid (though also may be other shapes), length greater than 4mm less than 1m, comparatively commonly greater than 4mm less than 600mm, more commonly less than 400mm; Width greater than 4mm less than 1m, comparatively commonly less than 500mm, more commonly less than 400mm; Thickness greater than 0.01mm less than 5.0mm, comparatively commonly greater than 0.1mm less than 2mm, more commonly greater than 0.2mm less than 1mm.When substrate 10 when being flexible, it can have all lengths, comprises 1m at least, 2m at least, or 5m (even 10m) at least at least.For the array of reading by detection fluorescence, substrate 10 can be made by the material that can launch slight fluorescence when with excitation light irradiation.In addition, in this case, too slow if focussed laser beam is propagated in certain zone, substrate can be transparent relatively so, so that reduce the absorption of incident irradiating laser and heating subsequently.For example, the irradiates light that incides upper surface that substrate 10 can transmission at least 20% or 50% (even at least 70%, 90% or 95%), the whole integration spectrum that this can be by this type of incident light or detect at 532nm and 633nm place and to measure out.

When array 12 is that in-situ method by routine forms; perhaps by depositing the part that before obtained (as previously mentioned; when depositing the part that had before obtained; for example by using pulse sprinklers such as ink gun; deposition one droplet reagent is to form each feature in each circulation) and when forming; zone 17 can exist usually between feature, and they do not carry any polynucleotide.Should be appreciated that zone 17 can have various size and shape between feature.Each feature has all been carried predetermined polynucleotide (also may be the mixture of polynucleotide).With usual the same, A, C, G, T represent conventional Nucleotide." Link " (seeing Fig. 3 for details) representative and upper surface and first Nucleotide is covalently bound to link agent (molecule), " Cap " representative not with Nucleotide bonded molecule, will further describe this below.

Substrate 10 also can comprise the identifier of one or more barcode 356 forms.For example also can use other the light or the identifier of magnetic identifier to replace barcode 356, the information of being discussed below it will carry.Because each identifier and each array 12 are positioned on the same substrate 10, and therefore has the fixed position with respect to barcode 356, can discern each array by described relative position, in this way, each identifier and each array 12 are linked together.Substrate can also have one or more benchmark sign 18, so that calibrate in the preparation of array and reading process.

Fig. 2 and Fig. 3 show the desired characteristics 16 of array 12a, 12b, and wherein formed actual characteristic is identical with target (or " purpose ") feature, and the shape of each feature 16, size and composition all are homogeneous, and these features have the interval of rule.When being prepared by the drop sedimentation, a such array all is the shape homogeneous with all reagent droplet that require to be used for each feature, and accurately is deposited on the target signature site.In practice, because systematic error and random error in the manufacturing processed, such desired result is actually and be difficult to obtains.For convenience of explanation, the feature 16 among the shown array 12a distributes in rectangular arranged mode (that is to say, rule row and column), and the feature 16 among the array 12b then distributes in hexagonal arrangement mode closely.In practice, all arrays 12 that are positioned on the same substrate 10 will use essentially identical decoration form (rectangle or tight hexagon).

One or more arrays with same characteristic features can be by recasting on surperficial 11a, and wherein, the recasting array has identical or different feature probe density.In this embodiment, all features that are positioned at an array have identical feature probe density, and probe combines with surperficial 11a by the link agent that is called as " Link " in Fig. 3.On each of feature 12a-12c, also exist and add cap agent (" Cap ").The suitable cap agent that adds can specifically be first silane that has carried out detailed description among any US 6,444,268, and the link agent can be any second silane wherein, and solvent also can be (for example toluene) described in this patent.As mentioned above, by reference this patent is included in herein herein, comprises the details of wherein employed first silane, second silane and solvent.In one embodiment, described in above-mentioned patent, first silane has formula R ¹-Si (R ^LR ^xR ^y), second silane has structural formula (before being linked on the biomonomer that is deposited) R ²-(L) _n-Si (R ^LR ^xR ^y), thereby make and to provide thereon-Si-R with the combining of surface ¹Group and-Si-(L) _n-R ²Group, wherein R ^LPart is a leavings group, can be identical or different; R ^xAnd R ^yBe low alkyl group or leavings group independently; R ¹It is the chemically inert part that behind the bonding to substrates surface, reduces its surface energy; N is 0 or 1; L is the link group; R ²Be make its can with the covalently bound functional group of molecular moiety, but perhaps can be converted into the modification group of this type of functional group.Above-mentioned leavings group can comprise halogen and alkoxyl group.The active hydrophilic part that first and second silane all pass through on it links to each other with the surface, and described hydrophilic segment is selected from the group that is made of hydroxyl, carboxyl, sulfydryl, amino and combination thereof.In above-mentioned patent, also above-mentioned term and other embodiment to first and second silane carried out further qualification.Substrate 10 can be handled with the above-mentioned silane mixture that describes in detail among the US 6,444,268, obtaining the second hydroxy-end capped silane group, the Nucleotide phosphoramidite can with this radical reaction so that link to each other with the surface by second silane (connection among Fig. 3).This has produced the decoration form shown in Fig. 1-3, and wherein, polynucleotide (or other biological macromole) links to each other with substrate surface 11a by the link molecule at feature 16 places.Can regulate the relative quantity of first and second silane, so that control surface can (thereby control hydrophobicity), US 6,444, also this described in detail in 268.

As can be seen from Figure 1, the successive zone of substrate 10 upper surface 11a is continual physically, and wherein upper surface 11a comprises surf zone 14 and array 12 between array.Therefore, surf zone 14 itself also is continual physically between array 12 and array.

Owing to all there is polynucleotide (having its hydrophilic functional groups) on each feature 16, so zone 14 is compared forr a short time between the hydrophobicity of each feature 16 and array, and and then makes the hydrophobicity of each array 12 also than regional 14 littler between array.Therefore, each array 12 is all surrounded effectively by the discontinuous of surface energy, and the discontinuous of surface energy can be so that the liquid sample that separates that is applied on the array 12 keeps separate stage.Compare with the zone that hydrophobicity is littler, the contact angle of zone that hydrophobicity is bigger and water-based drop is bigger, and therefore, the liquid sample that is applied on each array 12 tends to be limited on that array.This effect can be observed on array element 15, as shown in Figure 5.Fig. 5 has roughly described the relation of contact angle and droplet size on such as the surface of surperficial 11a.Increase along with the droplet size on the array 12 (its hydrophobicity is littler than zone 14 between array), as long as the surperficial contact area of drop remains in the middle of the array, the average contact angle of drop will roughly maintain on the constant so, is about 20 to 40 degree (having discontinuous between feature 16).But, when drop continue to increase, touching between the stronger array of hydrophobicity at regional 14 o'clock, contact angle begins to increase (shown in the near zone of vertical dotted line); When the surperficial contact area of drop enters between array regionally 14 the time, contact angle finally is stabilized on a certain value again.Under Fig. 1-Fig. 3 and situation shown in Figure 5, consider the average contact angle of the drop on each array 12, in fact surface energy is discontinuous, so near the average contact angles each array 12 also are discontinuous.

Array element 15 of the present invention can prepare in the following way: at first according to US 6,444,268 described methods utilize first and second silane that all surperficial 11a is functionalized.Then, the preparation method by above-mentioned in-situ preparation method or the previous biomacromolecule that obtains of deposition forms feature 16, thus on surperficial 11a preparation array 12.This can be undertaken by following manner: having on a plurality of features site of each array on the substrate to be prepared 10, contain biomacromolecule or other chemical probes or probe precursor (biological example monomer to the functionalized area deposition of substrate surface successive, as nucleoside phosphoramidites) drop, thereby make probe or probe precursor the feature site with link agent and combine.This step can repeat at one or more features place, particularly when adopting in-situ method to prepare biomacromolecule, up to finishing array 12.At length disclose these steps in the following document: for example, US 6,242, and 266, US 6,232, and 072, US 6,180, and 351, US 6,171, and 797, US 6,323, and 043, the sequence number that people such as Caren submitted on April 30th, 1999 is 09/302,898 U.S. Patent application, and the document of wherein quoting.

Because the formation of feature, when polynucleotide at each feature locations overtime, compare with zone 17 between functionalized feature, the hydrophobicity of each feature 16 (thereby each array 12) becomes littler.This is owing to have hydrophilic functional groups on the polynucleotide (though also can use other biomacromolecule with hydrophilic radical, for example peptides) that is prolonging.When feature 16 enough closely is filled in the array 12, though the zone 17 is still keeping their unreacted first and second silane between feature, and regional 14 to compare between array, the integral surface characteristic of array 12 hydrophobicity that will become is littler.A kind of simple method that obtains the required characteristic density of this effect of assessing is, with the identical functionalized surfaces of final array of expection (for example aforesaid undertaken functionalized surperficial 11a) by first and second silane on, makes up some but hot-wire arrays that characteristic density different identical with the final array composition of expecting.Detect the performance of water-based drop (preferably having identical composition) when volume increases that is positioned on the hot-wire array then with the sample that the array of reality will contact later.If observed behavior shown in Figure 5, make if drop do not reach liquid will be by crowded maximum volume to zone between array, the drop that is placed on the hot-wire array will often be limited, this density is exactly enough so.The region intermediate rising ground of the figure line of Fig. 5 is fast more steep more then good more.Also can use other equipment, to detect the resistance that drop moves when exceeding array 12.

Array element of the present invention can be by using any manufacturing the in the device described in the following United States Patent (USP): 6,420,180; 6,323,043; 6,180,351.According to the working method of installing described in those patents, use as passing through of having described of front and before whole surperficial 11a have been carried out functionalized substrate 10 and prepare array element 15 with first and second silane.Specifically, the treater of device is controlled the preparation of array by original position or deposition method according to deposited picture, so that generate array 12 at substrate 10 (can cut out from bigger substrate).In each circulation, deposit a group reagent drop at each target signature 16 place, thereby make probe or probe precursor combine with different zone by the link agent.Among the preparation technology, in each circulation of technology, repeat monomeric deposition in position at each feature place.In needs, treater also can be delivered to substrate the submergence station and carry out intercycle step or final step, and this all carries out according to above-mentioned traditional original position polynucleotide array preparation.At any point place of operation in the described device process, treater can be with US 6,180, and the mode described in 351 makes each array be associated with identifier such as barcode 356.Identifier can carry and close the layout information that is positioned at the array 12 on the substrate 10, also can carry other information (for example about the information of the sample volume of each array 12 that can be applied to specific array element 15, comprise following one or more: the sample volume of recommendation, minimum volume and maximum volume).The typical maximum volume that can be applied to each array 12 is no more than 1000 microlitres, perhaps is no more than: 500,200,100,50 or 10 microlitres.Perhaps, identifier 356 can be linked with the file that carries any above-mentioned information, thereby makes and can obtain this document based on identifier.This document and can be stored in the fixed storer by the treater of manufacturing installation with linking of identifier, perhaps can be written in the portable storage media, then this medium is placed in the packing identical, so that be transported to the remote client place with corresponding array component 15.US 6,180, described other purposes of identifier in 351.

Should recognize that under the guaranteed situation of surface energy saltation zone of aforesaid encirclement array 12, array element 15 can be made by other modes, for example passes through photoetching technique.

When the client receives array element 15, may utilize manually or automatic gear, with itself and sample effect, described subscriber station is positioned at the long-range place of array manufacturing station at subscriber station.Fig. 6 shows a kind of automatic gear.Treater 414 can receive any code (for example barcode 356) that reads from array element 15 from reader 400.Treater 414 also can be observed benchmark sign 18 by photographic camera 404, checks the aligning of substrate unit 15 with respect to sample allocation units 406.Sample allocation units 406 can be pulse sprinklers, and its structure is similar to 410, and can be piezoelectricity or thermal pulse shower nozzle, so that sample is sent to array 12 (with such as US 6,221,653 described modes).Perhaps, sample allocation units 406 can be automatic liquor-transferring systems.X-Y translate stage 408 can be with 406 an any position that move on the surperficial 11a.Treater 414 can also be by communication module 410 visit communication passages 180.Communication passage 180 can be any suitable communication passage, for example LAN, WAN, satellite or telephone network.

In operation, array element 15 manually is placed on 406 below, and treater 414 utilizes photographic camera 404 and benchmark sign 18 to determine that array elements 15 are with respect to 408 position.Reader 400 reads code (for example barcode 356), and obtains the layout of array 12 and about the information (for example following any one or multiple: the sample volume of recommendation, minimum volume or maximum volume) of each array 12 sample volume.The information of obtaining can be from the code that is read itself, from local storage, and with the form of the file that is linked with code this type of information is stored in this local storage in advance, perhaps from the remote memory that contains chained file (for example being positioned at array manufacturing works or elsewhere).Then, the information Control translate stage 408 that treater 414 utilization is obtained and 406, so that with sample deposition to each array 12, wherein sedimentary volume is to recommend volume on each array, at least be minimum volume, perhaps be no more than maximum volume, this depends on that the information that is obtained is aforesaid any.

In the non-automatic system that is used for using array element 15, can save photographic camera 404,406 and translate stage 408.As an alternative, the user can manually transmit the code 356 that is positioned at the array element 15 of reader 400 belows, and treater can obtain and the aforesaid volume information of demonstration on watch-dog 410.Then, the user can treat that sedimentary volume manually pipettes on the array 12 with aforementioned.

No matter what adopt during deposited samples is above-mentioned automatic technique or manual technique, all isolating liquid sample is applied on each array 12, wherein, sedimentary sample volume is enough to cover this array 12 on each array 12, is no more than the maximum volume that this array can hold simultaneously and can extend into zone 14 between array.Therefore, isolating liquid sample will be present on the substrate surface 11a simultaneously, and can not extend array 12 and enter zone 14 between the stronger array of hydrophobicity.As mentioned above, zone 14 has been kept institute and has been applied separation between the sample between the stronger array of hydrophobicity, and moves between array in regional 14 by suppressing sample, and they are limited in their zone of array 12.This situation is illustrated in Fig. 7, and wherein, because the effect in zone 14 between the stronger array of hydrophobicity, sedimentary isolating sample 22a, 22b, 22c are kept separate stage, and is prevented from being expanded to outside their corresponding array 12a, 12b, the 12c.Note contact angle θ ₁Can allow sample 22 and each array 12 to keep in touch at least 1 hour, 4 hours, 6 hours or at least 10 hours.The difference of regional contact angle can be greater than 10,20,30,40,60,70 or 80 degree between the average contact angle of array and array.

When the probe on the feature 16 and when to remain with its bonded sample all be polynucleotide to be hybridized, can use known hybridization array condition.In crossover process, array element 15 can be maintained in the wet environment (for example less sealing chamber of volume), perhaps can add the composition such as wetting Agent for Printing Inks in every kind of sample, and it can strengthen the maintenance of moisture.Wetting Agent for Printing Inks comprises: polyoxyethylene glycol (for example molecular-weight average 500,1000 or bigger polyoxyethylene glycol), triacetin, Sorbitol Powder, poly-dextrose, maltose alcohol, glycerine or tetrahydroxybutane, and promote other polymkeric substance (for example hydrophilic polymer) or the molecule that moisture keeps.Certainly, selected any specific examples of such components in other respects all should be compatible with the chemical property of hybridization solution and array.Also can in crossover process, adopt the volume (increase concentration) of controlled evaporation to reduce sample.Under the sort of situation, people may wish to adopt rigorous damping fluid at initial period, because along with the reducing of volume, preciseness can reduce.Perhaps, by in crossover process, adding hybridization buffer, also can obtain reverse effect.

Notice that sample size required when all being evenly distributed on surperficial 11a upward only as an array with the feature of all arrays is compared, in any case required sample volume all can reduce.This is because a big chunk of load characteristic surface 11a (zone 14 between array) does not contact with sample.Therefore, required sample volume can reduce, and perhaps employed sample concentration can be higher to promote hybridization.And the zone that the relative wetting ability of load characteristic is stronger helps still to guarantee that the sample uniform spreading is on array 12.

Sample effect, can washed with dry so that read it after the sufficiently long time in array.Notice that therefore all arrays 12 can, be compared with the situation that each array all is positioned on the substrates of different simultaneously with same dcq buffer liquid flushing, the otherness that this has reduced technology has improved processing power.

All arrays 12 on the unit 15 can be read simultaneously by any suitable reading device.When in target, having introduced fluorescent mark with known approaches thereby fluorescence to be detected is arranged, can use known array reader.For example, this type of reader can be in the mode of grating at one or more illuminating laser beams of entire area interscan of each array and the fluorescence of detected any generation, as US 6,406, described in 849.Note, array element of the present invention can avoid using any extra such as wax hydrophobic material or to the demand of liner etc., and all these may introduce a spot of fluorescence pollutent.Since very faint from the fluorescent signal that each array features read, therefore, even the concentration of this pollutant is very little and only in some site existence, also might have a strong impact on the result.

The reading result of array can be treated result, for example obtain by following manner: refusal is to a certain reading that is lower than the feature of predetermined threshold, and/or form conclusion (for example whether may have a kind of concrete target sequence the sample, perhaps whether the organism of sampling has shown certain specific patient's condition or disease) based on the pattern that reads from array.The result who reads (handle or be untreated) can pass through communication passage 180 or reader/write device 186 and medium 190, is forwarded (for example by communication) to remote location and receives, so that further assess and/or handle or use.If desired, can transmit with the arrival remote location these data by other means, or be transported to other positions as required again.

In a variation of the foregoing description, each array component 15 might be included in the suitable outer cover.This type of outer cover can comprise usually and can conduct interviews to it by one or more ports, and described outer cover to be carried substrate 10 by the chamber of the sealing that partition sealed.In this case, being positioned at the identifier of all arrays on the substrate 10 can be by being applied on the described outer cover and be associated with them.Also should recognize, can read array by any other method or device that the front is not mentioned, wherein, other reading method comprises that other optical technology (for example detecting chemoluminescence or electroluminescence mark) or electroporation (wherein provide electrode to each feature, to detect the hybridization at this feature place, its mode such as US 6,251,685, US 6,221,583 etc. disclosed).For from feature picked up signal data (feature extraction), wherein feature and corresponding signal thereof are identified in the array image that is read in this operation, can carry out according to the process of describing in the following patent: sequence number is 09/589046,09/659415 and 10/086839 U.S. Patent application, its title are " Method And System For Extracting Data From Surface ArrayDeposited Features ".

In the shown embodiment of Fig. 1-3, each regional border 19 of the carrying array 12a that surrounded by zone 14 between array (being defined by the line on the outermost border of contact array 12a outermost feature 16 just) provides the surface energy saltation zone, to keep the separation between the isolating liquid sample that is applied on the array.But this is not essential.For example, between array and such surface energy saltation zone, a gap can be arranged, wherein, provide other components littler (then surface energy saltation zone between the stronger array of this composition and hydrophobicity between the zone 14) than array 12a hydrophobicity at this gap location.In another alternative embodiment, array needs not to be even layout.For example, as shown in Figure 8, the external region 22 (also the border 19 with array 12a is the boundary) that array 12a can comprise interior region 20 (is the boundary with 21) and surround interior region 20.External region 22 also can have a plurality of features 24, and these features 24 can all have identical or different composition (if the composition that they contain is stronger than the wetting ability in zone 14 between array).For example, feature 24 can be the polynucleotide with identical sequence.The ratio of feature 24 shared external region areas (being limited between boundary line 21 and the border 90) is bigger than the ratio of feature 16 shared interior region areas (is the boundary with boundary line 21).Therefore, in fact the wetting ability of external region 22 may be bigger than interior region 20.Notice that if external region 22 is filled continuously by the component institute that wetting ability is higher than zone 14 between array, this will represent an embodiment so, and a gap is wherein arranged, as previously described between array and surface energy saltation zone.In this case, the border of array 12a is 21 places in the boundary line, and 19 is the surface energy saltation zone.Like this, in this embodiment, the hydrophobicity in zone 14 is stronger than the border 19 in besieged zone between array.

The substrate surface that polymerized nucleoside acid constituents or other parts will deposit thereon can be a porous or non-porous, can be level and smooth or is plane substantially.Substrate can be a kind of material or have multilayered structure.When wishing that array has certain pattern, in Fig. 1, the regular row-column configuration of array 12, can also construct various geometrical shapies.For example, array 12 can be arranged to row (point of for example a series of concentric circles or semicircle shape) of a series of curves that are covered with substrate surface or the like.Similarly, the pattern that is arranged in the feature 16 of array 12 can be different with the well-regulated ranks shape of Fig. 2 feature, can comprise for example row of a series of curves (for example a series of concentric(al) circless or semicircle) or the like.Similarly, the pattern of the feature 16 in array 12 can be different from the row or column of the several rules of the feature among Fig. 2, and the pattern of feature 16 can comprise for example row of a series of curve (for example, for example a series of concentric(al) circless or semicircular point) or the like.Though can adopt various feature placement modes, the mode (for example by the array identifier) of some some characteristic at least that can confirm feature (for example following is any or multiple: the component of feature, site, size, with at the performance characteristic that provides in conjunction with the form of the significance of the variation of pattern with different samples etc.) should be provided to the user.Can select the decoration form of array according to the consideration of manufacturing, processing and use aspect.Present method and the device can with aforesaid similar mode, be used to make and use the array that has other biological macromole, polymkeric substance or other parts on the surface, its mode is with foregoing identical.Correspondingly, when mentioning polymkeric substance, biomacromolecule or polynucleotide etc., often can replace with " chemical part ".

Certainly above-mentioned specific embodiment is carried out various further modifications.Therefore, the present invention's specific embodiment of being not limited to describe in detail previously.

Claims (24)

1. array element, comprise the substrate surface that is carrying a plurality of biomacromolecule arrays, all comprise a plurality of features in each array, and each array is all surrounded by the surface energy saltation zone, keeping the separation between the isolating liquid sample that is applied on the described array, and wherein said substrate surface is physically continual on the successive zone of the described array of carrying.

2. array element as claimed in claim 1, wherein, described surface energy saltation zone is provided by surf zone between array, and surf zone is between the zone of carrying array between described array, and it is surround the zone of described carrying array, and stronger than the hydrophobicity of besieged zone iimit.

3. array element as claimed in claim 2, wherein, the border in described besieged zone is the border of described array.

4. array element as claimed in claim 2, wherein, described biomacromolecule comprises polynucleotide or peptide.

5. array element as claimed in claim 4, wherein, surf zone is stronger than the hydrophobicity of described array features between described array.

6. array element as claimed in claim 5, wherein, between the described array before the zone, the volume of the water that described array can hold can reach V at water intrusion, and wherein V is provided by following formula:

$V(r,{\mathrm{\θ}}_c)=\frac{1}{3}\mathrm{\π}{\left(\frac{r}{\cos (\frac{\mathrm{\π}}{2}-{\mathrm{\θ}}_c)}\right)}^{\frac{1}{3}}(2-3\sin (\frac{\mathrm{\π}}{2}-{\mathrm{\θ}}_c)+\sin {(\frac{\mathrm{\π}}{2}-{\mathrm{\θ}}_c)}^3)$

Wherein, V (r, θ _c) be the volume of sessile drops; R is the radius of circular array, and perhaps when array is not circle, r is the radius of the circle that equates with described array area; θ _cIt is contact angle greater than 20 degree.

7. array element as claimed in claim 5, wherein, the contact angle of described array features is less than 20 degree, and the feature total area accounts at least 30% of each array area.

8. array element as claimed in claim 4, wherein, described array is circular.

9. array element as claimed in claim 4, wherein, the minimum interval between the described adjacent array is at least two times of mean distance between the described array features.

10. array element as claimed in claim 5, wherein, the minimum interval between the described adjacent array is not more than ten times of mean distance between the described array features.

11. array element as claimed in claim 4, wherein, described each array comprises 50 to 1000 features.

12. array element as claimed in claim 4, wherein, surf zone comprises the dissimilar molecule that is connected with described substrate surface between described array.

13. array element as claimed in claim 12, wherein, each feature of described array also comprises the dissimilar molecule that is connected with described substrate surface, wherein has only a subclass of described dissimilar molecules biomacromolecule can be covalently bound on the inner surface of described feature.

14. array element as claimed in claim 4, wherein, each array comprises interior region and surrounds the external region of described interior region, a plurality of features are contained in each zone, and the ratio of the described interior region area that the ratio of the described external region area that feature is occupied is more occupied than feature is bigger.

15. array element as claimed in claim 14, wherein, the described external region of each array comprises at least eight features.

16. a method of using the described array of claim 4 comprises the following steps:

On each array, apply isolating liquid sample, thereby make a plurality of same times of isolating sample be present in described substrate surface, simultaneously between the array that hydrophobicity is stronger zone maintenance the separation of the sample room that applies.

17. method as claimed in claim 16 wherein, does not have physical barrier between the array.

18. method as claimed in claim 16 wherein, comprises polynucleotide or peptide, and comprises polynucleotide or peptide in each sample in the described array features.

19. a method for preparing the described array of claim 4 comprises the following steps:

(a) drop that will contain the biomacromolecule precursor deposits to the site that feature to be formed is arranged on the described substrate surface, thereby makes described precursor combine with described surface in described site;

(b) repeat (a) step in each described site, previous bonded precursor is combined with the precursor that is deposited subsequently, be produced out up to described array;

Wherein, after repeating (a) step, compare with zone between described array, the regional hydrophobicity of described formation array becomes littler.

20. method as claimed in claim 19, wherein, described substrate surface comprises the composition that combines with described substrate surface, and described composition had both covered zone between described array, also covers the zone of the described formation array that comprises described feature site; In step (a), at least a composition in the described composition is attached to described biomacromolecule precursor on the described surface.

21. method as claimed in claim 20, wherein, described composition comprises the dissimilar molecule that combines with described substrate surface, but has only a subclass of described dissimilar molecules that biomacromolecule is covalently bound on the described surface of described feature inside.

22. a use comprises the method for the array element that is positioned at a plurality of biomacromolecule arrays on the substrate surface, each array all comprises a plurality of features, and described array is separated by surf zone between array, wherein, described substrate surface is physically continual on the successive zone of carrying array, and described method comprises the following steps:

Apply isolating liquid sample to each array, thereby make a plurality of same times of isolating sample be present in described substrate surface, keep separating by zone between described array simultaneously.

23. method of using the described array element of claim 4, described method comprises the following steps: to read the code that is associated with described array element, and from described code or with file that described code is linked obtain the relevant information that should be applied to the liquid sample volume on the described array.

24. method as claimed in claim 23, wherein, described liquid sample is an aqueous sample, comprising wetting Agent for Printing Inks to keep moisture.

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